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Plasmide DNA isolation from bacteria and transfection to HEK293 cell line
Skala, Kateřina ; Fohlerová, Zdenka (referee) ; Svoboda, Ondřej (advisor)
DNA isolation is one of the basic methods in molecular biology. There are several methods of DNA amplification and isolation. In this paper phenol-chloroform extraction of three plasmid types - Channelrhodopsin-2, ASAP1 and Kir 2.1 is used. Six plasmids were isolated in total. These plasmids are then validated using gel electrophoresis. Successfully isolated plasmids are then transfected to HEK293 cells and images taken on confocal microscope 24, 48 and 72 hours after transfection.
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Project of revitalization part of Kninicky brook
Vyplel, Adam ; Svoboda, Ondřej (referee) ; Šlezingr, Miloslav (advisor)
The diploma thesis is focused on the proposal of systematic revitalization of the selected section of the Knínický brook in the cadastral area of Veverské Knínice. It is a straight, very deep channel due to technical adjustments in the past. The HEC-RAS program verifies the capacity of the channel for the Q100 flow. In this way, it was found that the channel has very high capacity so a new route with Q1 flow capacity was designed. Due to the height conditions, the design also included three boulder chutes. Subsequently, the vegetation accompaniment and bank stands were suitably supplemented.
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3D bioprinting of stem cells and analysis of microscopic images
Kandra, Mário ; Svoboda, Ondřej (referee) ; Kolářová, Jana (advisor)
In this diploma thesis we are discussing about using 3D bioprinting in tissue engineering. We are discribing using biomaterials for construction scaffolder and aplication stem cells in 3D bioprinting. Last section of theoretical part deals with very often used techniques of 3D bioprinting and we are focused on extrusion technique. In the practical part we propose a method for print vasculars structures. We realized prototype of print head, her design and 3D printing of individual parts. To mechanical part we create a control system for printing control. At the end we visualize the organization of the cells using program modules.
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Study of properties of voltage membrane sensor ASAP1 expressed in HEK293 cell line
Sanetrníková, Dominika ; Chmelíková, Larisa (referee) ; Svoboda, Ondřej (advisor)
In the beginning of this thesis is a short introduction into plasmid DNA which is in the form of a vector used in molecular biology. Plasmids can be used in the form of fluorescent probes to measure changes in membrane potential. Into their structure is added a dye called fluorophore. As an important representative of this thesis is a fluorescent probe ASAP1 which contains green fluorescent protein whose response to the membrane potential change is the decrease in the intensity of emitted light. The aim of this thesis was to make chemical transfection of this plasmid into the HEK293 cell line and carry out its characterization. In the work is also described the design of a method for the analysis of the time course of changes in fluorescence depending on the cell membrane depolarisation. In the end of this thesis is also desribed realized experiment including the discussion of aquired results.
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Study of cardiomyocytes viability using confocal microscopy
Chutný, Pavel ; Svoboda, Ondřej (referee) ; Čmiel, Vratislav (advisor)
This work describes the study of cardiomyocytes viability using confocal microscopy. The theoretic introduction covers the basic principle of fluorescence together with fluoresceins in cells biology. The struction of cardiomyocytes, their selection from the lively heart and the methods of viability examination is comprised in the next part. The third topic is about confocal mikroskopy and its posibilities in cardiomyocytes imaging. In next chapters are described the experiments and their evaluating.
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Evaluation of Automatic Vessel Tree Segmentation Algorithms
Svoboda, Ondřej ; Jan, Jiří (referee) ; Odstrčilík, Jan (advisor)
Segmentation of the vasculature is an important step in the process of the retinal image analysis. The results of the analysis can be used to diagnose several eye and cardiovascular diseases. This work deals with the creation of gold standard database of high resolution retinal images and their use in evaluating the success of vascular automatic segmentation methods. The aim is to create the application, which will online evaluate the success of the automatic vessel segmentation methods. In brief, this work describes the characteristics of image data from digital fundus camera, the method of image segmentation and automatic segmentation methods of blood vessels. Furthermore, this work describes the gold standard, the databases of gold standards and ultimately the properties of the new database and the reason for HRF (High Resolution Fundus Images). The last chapter deals with methods of evaluating the success of vascular automatic detection methods and application development for this assessment.
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Magnetic Nanoparticles for Targeted Delivery System of Plasmid into HEK293 cells
Bílek, Ondřej ; Svoboda, Ondřej (referee) ; Fohlerová, Zdenka (advisor)
This bachelor’s thesis deals with using of magnetic nanoparticles, SPIO nanoparticles and nanoparticles of commercial product MATra for transfection of ASAP plasmid into HEK293 cells and subsequent optimization of the process. The experimental part is enclosed with theorethical part that summarizes findings from the fields of synthesis of magnetic nanoparticles and their biomedical applications, theory dealing with possibilities of the insertion of foreign DNA or plasmids into the cell and of its efficiency evaluation.
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Application of fluorescence staining for the "Scratch Wound Healing Assay" experiment
Zumberg, Inna ; Svoboda, Ondřej (referee) ; Chmelíková, Larisa (advisor)
This bachelor’s thesis studies the cell culture of 3T3 cell line (mouse embryonic fibroblast cells) and cell migration. The Scratch Wound Healing Assay was used for the study of cell migration. The basic principles of experimenting, its advantages and disadvantages, as well as the practical use of this method in scientific research were presented on specific examples. Practical part of the bachelor’s thesis describes the experiment in the cell laboratory including cell culture of 3T3 cell line. The proposed experiment was tested with a sufficient number of repetitions. The statistical processing of microscopic images was performed in the MATLAB programming environment.
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Proposal of Implementation of Business Strategy with IS/IT Support
Svoboda, Ondřej ; Polák, Vojtěch (referee) ; Zich, Robert (advisor)
This master's thesis deals with a implementation of IT/IS tools to support the implementation of the company's business strategy, which sells and provides after sales servicematerial handling equipment. The thesis examines key internal and external factors related to the implementation of business strategy of the company, evaluates them and proposes concrete implementation of IT/IS tools to support achievement of strategic goals.
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